Extended Data Fig. 2: ECSIT promotes the recall response of memory CD8⁺ T cells.

(a) Schematic of memory CD8⁺ T cells recall response. Naive CD8⁺ T cells obtained from Ecsit^fl/fl OT-1 or Ecsit^fl/fl dLck-Cre OT-1 mice were transferred into WT recipient mice. Subsequently, the recipient mice were infected with VSV-OVA 1 day later. After 35 days, memory OT-1 T cells were sorted and transferred into naive WT recipient mice, followed by LM-OVA challenge. Spleen and liver were collected for analyses 5 days later. (b, e) Representative flow cytometry plots (top) and the analysis of percentage and number (bottom) of Kb-OVA⁺ CD44⁺ CD8⁺ cells in the spleen (b) and liver (e). (c, d, f, g) Representative flow cytometry plots (top) and the analysis of percentage and number (bottom) of IFN-γ⁺ (c) and TNF-α⁺ (d) T cells gated on Kb-OVA⁺ CD44⁺ CD8⁺ cells in the spleen, and IFN-γ⁺ (f) and TNF-α⁺ (g) T cells gated on Kb-OVA⁺ CD44⁺ CD8⁺ cells in the liver. (h, i) Bacterial load of LM-OVA in the spleen (h) and liver (i). (j) Schematic of memory T cells formation in acute infection. Naive CD8⁺ T cells from Ecsit^fl/fl OT-1 or Ecsit^fl/fl Rosa26^CreERT2 OT-1 mice were transferred into WT recipient mice, followed by VSV-OVA infection 1 day later. Tamoxifen was intraperitoneally injected from day21 to day25. OT-1 memory cells were analysed on day35. (k) Schematic of memory T cells formation in acute infection with adaptive co-transplantation model. Naive CD8⁺ T cells from Ecsit^fl/+ CD45.1.2 OT-1 and Ecsit^fl/fl Rosa26^CreERT2 CD45.2 OT-1 mice were mixed equally and transferred into CD45.1 recipient mice, and then the recipient mice were infected with VSV-OVA 1 day later. Tamoxifen was intraperitoneally injected from day21 to day25. OT-1 memory cells were analysed on day35. Data are pooled from three independent experiments and n = 5 for (b-i). Error bars show mean ± sem. Two-tailed unpaired student’s t-test for (b-i).

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